JP2009536931A - Novel use of 4,17β-dihydroxyandrost-4-en-3-one - Google Patents

Abstract

The present invention relates to novel uses of 4,17²-dihydroxyandrost-4-ene-3-one (hereinafter 4-hydroxytestosterone), to a process for their preparation, to pharmaceutical compositions containing them, and to the use of said compounds for the prophylaxis and/or treatment of hormone-dependent or hormone receptor regulated cancers in mammals.

Description

  The present invention includes 4,17β-dihydroxyandrost-4-en-3-one (hereinafter referred to as 4-hydroxytestosterone) and novel uses of these salts and esters, methods for their preparation, and the like The present invention relates to pharmaceutical compositions and the use of these compounds in the prevention and treatment of certain diseases in mammals, in particular carcinomas.

  Breast cancer is mostly related only to women, but after colorectal cancer, breast cancer is the most common cancer disease in Western countries. In Germany, breast cancer accounts for about 20% of all cancer types diagnosed in women. The treatment of most cases consists of surgical removal of the tumor (mastectomy of the adenocarcinoma) or complete removal of the affected breast if the size of the tumor interferes with breast conservation therapy. In both cases, drug therapy continues after surgery. A particularly troublesome problem with breast cancer is its high ability to metastasize to other organs, particularly the liver, brain, bones, and skin. Surgery therefore involves drug therapy (adjuvant therapy). This therapy is also aimed at preventing local recurrence of the disease.

If the removed tumor expresses an estrogen receptor, an anti-estrogen is used (Tamoxifen or an aromatase inhibitor). If the estrogen receptor is missing, surgery with a cytostatic reagent It will be post-chemotherapy.
Tamoxifen binds to the estrogen receptor (ER) and inhibits the growth-promoting effect of estrogen. Aromatase inhibitors inhibit the final stage of estrogen production from androgen precursor (C19-steroid). These bind irreversibly to the active site of the enzyme {aromatase-inactivating agent, ie steroidal aromatase inhibitor: 4-hydroxyandrostenedione (Formestan = Lentaron®) or exemestane (Exemestan (Aromasin ( R))), or the cytochrome p450 part of the enzyme {non-steroidal aromatase inhibitor: anastrozol (Animidex (R))) and lentrozol (Letrozol (Femara (R))} competitively.
If breast cancer overexpresses the Her2neu gene, an antibody against this cell surface protein, eg, Trastuzumab (Herceptin®), is administered intravenously.

  According to basic and clinical data, aromatized metabolites of androgens, ie estrogens, are pathogenic cellular changes associated with the growth of several hormone-dependent cancers such as breast cancer, intrauterine ovarian cancer It is understood that it is a hormone involved in. Aromatase (aromatizing enzyme) has recently been recognized as an enzyme capable of synthesizing estradiol directly in tumors.

  Endogenous estrogen is ultimately made from androstenedione or testosterone as a direct precursor. Local estrogen production in breast tissue, especially the tumor cells themselves, has been considered of particular importance for the treatment of this disease. In this context, the aromatization of steroid ring A performed by the enzyme aromatase and its inhibition has been thought to play a major role in anti-breast cancer methods.

  Prior to the present invention, apart from other concepts based on estrogen receptor antagonists or antibodies used for treatment, conventional treatment methods for addressing female breast cancer have therefore focused on the aromatase inhibitory effect itself. Accordingly, steroidal substances reported to have an aromatase inhibitory action are described, such as Δ1-testrolactone [US Pat. No. 2,744,120], 4-hydroxyandrost-4-ene-3,17-dione and esters thereof. [See, eg, US Pat. No. 4,235,893] 10- (1,2-propadienyl) -estro-4-ene-3,17-dione (US Pat. No. 4,289,762, 10- (2-propynyl) -estro-4-ene- 3,17 dione [J. Amer. Chem. Soc., 103, 3221 (1981) and US Pat. No. 4,322,416], 19-thioandrostene derivatives (European Patent Application No. 100566), Androsta-4,6-diene-3, 17-dione, androsta-1,4,6-triene-3,17-dione [UK Patent Application 2,100,601A] and androsta-1,4-diene-3,17-dione [Cancer Res. uppl.) 42, 3327 (1982)].

  However, according to clinical studies conducted using estrogen receptor (ER) antagonists and aromatase inhibitors, these compounds do not have significant and sufficient inhibitory effects on tumor growth and / or growth. I understood. For example, Tamoxifen (R) -based therapy suffered from disorders like tachyphylaxis (treatment failure) and the fact that some cells have estrogen-like effects (eg thrombosis and uterus) Due to the risk of endometrial cancer, tamoxifen (Tamoxifen®) cannot be administered for 3 to 5 years or longer. New drugs, such as aromatase inhibitors, can in principle be used only in menopausal women and are also effective only in ER-positive tumors.

  Stable transgenic cells expressing AR have been reported to acquire the ability to respond to androgens and cause termination of cell proliferation (J. Szelei et al., “Androgen-Induced Inhibition of Proliferation in Human Breast Cancer MCF7 Cells Transfected. with Androgen Receptor ", Endocrinol., 138, No. 4, 1406-1412, 1997). According to a more recent report (J. Ortmann et al., “Testosterone and 5α-dihydrotestosterone inhibit in vitro growth of human breast cancer cell lines”, Gynecol. Endocrinol. 2002; 16: 113-120), testosterone and 5α-dihydrotestosterone ( DHT) inhibits cell proliferation in certain cell lines. Testosterone itself has a similar effect. However, this clinical efficacy is hampered by the fact that this compound is rapidly metabolized to estradiol or dihydrotestosterone (DHT). Local production of estradiol from testosterone by local aromatase activity in breast cancer tissue promotes cancer growth.

  Thus, other androgen receptors (AR) that exert testosterone-like effects (special anabolism) in AR positive cells and tissues and are not metabolized by aromatase or converted to DHT by 5-alpha-reductase. There is an increasing demand for specific compounds.

  According to the present invention, the therapeutic effect of 4,17β-dihydroxyandrost-4-en-3-one was discovered under a surprising new context, enabling valuable medical applications. That is, the present invention relates to 4,17β-dihydroxyandrost-4-en-3-one or a salt or ester thereof, breast cancer (particularly breast mucinous ductal carcinoma, medullary carcinoma of the breast and breast lobular carcinoma), intrauterine Membrane cancer, cervical cancer, epithelial ovarian cancer, uterine (fallopian tube) cancer, benign proliferative changes or alterations of the breast, fibroadenoma, micromastosis (perimenopause), mastopathy, uterine fibroids, and these diseases In the preparation of a pharmaceutical composition for the prevention or treatment of secondary or metastatic tumors.

  “Use” in the present invention may include treatment or prophylaxis of the specific compound, or a pharmaceutical composition comprising the compound as an active ingredient together with an appropriate carrier and / or diluent for the above indicated diseases. The term also includes use in the preparation of the pharmaceutical composition.

  Based on the findings in the present invention, 4,17β-dihydroxyandrost-4-en-3-one is particularly suitable for the prevention and / or treatment of breast cancer in female or male patients.

  The pharmaceutical composition of the present invention further comprises an effective amount of one or more estrogen receptor antagonists and / or one or more aromatase inhibitors (different from the compounds disclosed in the present invention) and / or one or more A cytostatic agent and / or one or more antibodies against Her2neu may be included.

  The pharmaceutical composition of the present invention is preferably used because the compound of the present invention can most effectively exert these effects directly on the target cells or tissues without systemically affecting the treated patient. Is formulated for topical treatment at the target location to be treated for local treatment.

  The compounds of the present invention exhibit androgen receptor (AR) binding affinity. In most cells except breast cancer cells, these cells can induce anabolism as long as they present AR. In breast cancer cells, these cells cause apoptosis. These compounds are not substrates of aromatase and are not metabolites and therefore do not share the negative effects of compounds metabolized by aromatase. Furthermore, the compounds of the present invention are not substrates for 5α-reductase. Since these compounds cannot be metabolized as easily as, for example, testosterone, the inhibitory effect of these compounds on tumor cells becomes very easy, and further shows the effect of causing apoptosis in the tumor cells themselves. . Thus, these compounds are probably more effective in the treatment of breast cancer cells than the series of compounds currently used to treat both types of breast cancer. Another beneficial effect of this prophylactic or therapeutic concept is that these cells are likely to be AR positive, even in estrogen receptor negative (ER-negative) tumors, and are therefore currently treated with growth inhibitory reagents alone. Tumors that can be treated pharmacologically with 4-hydroxytestosterone significantly increase the range of hormone therapy for breast cancer.

  Since the compounds of the present invention are AR specific compounds but are not metabolized by aromatase, these compounds exert actions and effects related to the specifically defined therapeutic or prophylactic use via AR. This makes the concept of the currently selected compound clearer than the prior art. This also explains the fundamental differences beyond the traditional therapeutic use of 4-hydroxytestosterone or its derivatives. In particular, the disclosure of US Pat. No. 2,762,818A is no more than the use of 4-hydroxytestosterone and its esters to treat androgen deficiency conditions themselves based on the androgenic and anabolic properties of these compounds. However, there has been no purpose or discovery to suggest an effect that can be used to deduce activity against breast cancer cells or its effectiveness against disease states according to the present invention. Furthermore, US Published Patent Application 2003 / 0229063A addresses the low androgen / estrogen ratio in men (endocrine disorders occur), and for this purpose, the use of 4-hydroxytestosterone is based solely on the claimed aromatase inhibitory effect. Try.

  In experiments conducted with the compounds of the present invention, these compounds have excellent skin permeability and, therefore, a therapeutically or prophylactically effective amount of the book on the part of the patient in need of treatment. It has been shown that simple topical administration, including, for example, ointments, topical aqueous solutions or creams, etc. containing the compounds of the invention is sufficiently effective on tumor growth (until apoptosis). After topical administration, the compound (s) penetrates the skin and is concentrated into subcutaneous adipose tissue.

  In the case of breast cancer, the drug can be applied topically to the breast skin, so that the active compound, which is preferably lipophilic, is absorbed transdermally and is therefore delivered locally to the intended site of action. Is done. The active compound collects in the periductal adipose tissue. In long-term treatment, the fat content of the treated breast is significantly reduced. This reduction reduces estrogen-forming cells with estrogenic performance. The lipophilicity and hydrophobicity of the active compound results in the active compound being extremely locally concentrated in the adipose tissue, and thus cannot exhibit any significant systemic effects.

According to a preferred embodiment of the invention, it is intended to apply the drug directly on or near the intended site of action. According to the present invention, the result obtained is that a sufficient local concentration of the active compound in dangerous tissue (in prevention) or diseased tissue (in treatment) without the active compound being significantly absorbed into the circulation. Indicates. The advantages of this embodiment are not only dependent on the local administration per se, but the active compound is directly accumulated in the dangerous and / or diseased tissue, not indirectly through the circulating blood stream, Depends on local application.
Especially in the case of breast cancer, since the primary target disease is treated as described above, female patients, especially female breast skin, are often applied locally or prophylactically. The same therapeutic ingredients are applied.

  If metastatic cancer is also treated and / or prophylaxis against metastatic cancer can be carried out, the agent of the present invention additionally has a significant absorption in the circulating bloodstream and thus the active compound is at the site of metastasis. Such an amount can be applied topically to the intended site of action so that serum levels are also transported. Also in this use, local absorption is first performed, especially on or within the intended site of action where metastatic cancer is present.

  An important advantage of the present invention is the possibility of using this drug and also for breast cancer prevention. A particularly important advantageous use possibility is the so-called secondary prevention. In female or male patients diagnosed and treated for breast cancer, there is a high risk of recurrence at the same site and a high risk of developing further cancer in the contralateral breast. In both cases, the occurrence of a secondary cancer with ER is unpredictable. The treated breast and the contralateral breast can be prophylactically treated with the medicament of the present invention. This could also prevent the development of ER negative cancers, especially common in patients with a familial predisposition to breast cancer.

Primary prevention can be performed in so-called high-risk women. The selection criteria used for such high risk groups are, for example, that at least one female or male relative, such as the mother's first parent, had breast cancer on one or both sides or had breast cancer before age 45 years. It is the fact that at least one female or male relative and a further male or female relative, such as a first parent, have breast cancer, or have had breast cancer, on the mother side.
In most of these cases, the underlying cause of the disease is a genetic deletion in BRCA1 and BRCA2 known as the two “BREAST CANCER GENES”.

In topical applications according to a preferred embodiment of the present invention, the compound's hydrophobicity and high safety profile virtually completely avoids the side effects of the active compound, so the primary prevention instruction is relatively low risk tissue. Or if there is a tissue with a moderate risk, it can be done relatively generously immediately. Prevention can begin long before the signs or symptoms of breast cancer occur.
In preferred topical administration, the compounds or compositions of the invention are preferably applied over a relatively long period of time (lifetime if necessary), and administration is performed, for example, once or twice a day. The active compounds according to the invention intended for topical (local) application are preferably provided in a fat-soluble form, so that they can easily penetrate adipose tissue and prevent them. Exert local and / or therapeutic effects locally.

  Active compounds administered topically according to the present invention remain localized to the adipose tissue of the breast and exhibit the intended effect by lipid solubility, thus eliminating any side effects caused by systemic application. The ability to reduce or eliminate this side effect allows for a very broad prophylactic use. The medicament of the present invention can be applied by the patient himself and frequent visits to the doctor are not necessary for this purpose.

  In general, for topical administration, the active compounds according to the invention are lipophilic and are very suitable for topical application. As already mentioned, accumulation in breast adipose tissue avoids systemic side effects. In order to improve skin permeability, compounds known in the prior art that promote this, such as hyaluronidase, dimethyl sorbitol or DMSO (dimethyl sulfoxide), can be added to the agents of the invention.

In particular, topical administration is primarily primary prevention; after breast-conserving treatment, along with various adjuvant drug treatments after surgery for 'in situ ductal carcinoma' (DCIS, usually not followed by any adjuvant treatment), Directed for secondary prevention; and for prevention of contralateral breast cancer.
Topical administration is also particularly suitable in the context of 'neoadjuvant' treatment. This means pre-operative cancer drug therapy to reduce breast cancer size for breast-conserving surgery. In the past, it has been mainly done for ER negative tumors, and recently aromatase inhibitors have been used successfully with restrictions within 6 months. Finally, treatment can be applied to superficial damage (eg skin metastases).

  Primary prevention relates to the prophylactic treatment of healthy individuals who are at risk of developing cancer. Since the compounds and pharmaceutical compositions of the present invention are not aromatase substrates, cannot be metabolized to estrogens, and do not maleize, they can also be used for primary prevention of breast cancer in premenopausal women. In secondary prevention, the compounds and compositions can be used alone or in combination with, for example, Tamoxifen, aromatase inhibitors, herceptin and / or cell growth inhibitors, such as Any of these combinations will expand the spectrum of cell types that can be treated.

  The present invention is based on the discovery that 4,17β-dihydroxyandrost-4-en-3-one (hereinafter referred to as 4-hydroxytestosterone) can be successfully used as a pharmaceutically active compound. In particular, a pharmaceutical composition comprising a suitable carrier and / or diluent and 4-hydroxytestosterone as an active ingredient is used in breast cancer (especially breast mucinous ductal carcinoma, medullary carcinoma of the breast and breast lobular carcinoma), intrauterine Membrane cancer, cervical cancer, epithelial ovarian cancer, uterine (fallopian tube) cancer, benign proliferative changes or alterations of the breast, fibroadenoma, micromastosis (perimenopause), mastopathy, uterine fibroids, and these diseases Can be used for the prevention and / or treatment of secondary or metastatic tumors.

  Accordingly, the present invention provides 4,17β-dihydroxyandrost-4-en- in the preparation of a pharmaceutical composition for the (clinical) prevention and / or treatment of the above hormone-dependent or hormone receptor-regulated cancer in mammals. The use of 3-one (4-hydroxytestosterone) is provided as well.

  In general, the compounds and pharmaceutical formulations of the invention may be administered topically, orally, rectally, parenterally (eg, intramuscular or intravenous injection, or infusion), transdermally and / or It can be administered as a sustained formulation.

  The compound of the present invention, 4-hydroxytestosterone, is disclosed, for example, in US Pat. No. 2,762,818A and is commercially available (eg, Bulk Nutrition, Graham, NC. USA; further information is bulknutrition.com WINKOS GmbH D-79189 Bad Krozingen, DE).

  The present invention also relates to all derivatives that share functional and / or structural characteristics (particularly salts and esters of 4,17β-dihydroxyandrost-4-en-3-one). Examples of suitable ester groups include straight, branched, or cyclic or aromatic acyl groups such as formyl, acetyl, propionyl, butyryl, pentanoyl, hexanoyl, heptanoyl, and benzoyl. There is, but is not limited to these. These esters can be made having 4 and / or 17 hydroxy groups, preferably 17 hydroxy groups. These salts and esters can be prepared by known methods (see, for example, US Pat. No. 2,762,818A).

  The compounds and pharmaceutical formulations of the present invention are anabolic steroids and exert testosterone-like effects by binding to the androgen receptor represented by AR positive cells and tissues.

  Given these high therapeutic indices, the compounds of the present invention can be used safely in medicine. For example, the approximate acute toxicity (LD50) to mice of a compound of the present invention measured in a single dose with increasing dose and measured 7 days after treatment is negligible.

  The compounds and pharmaceutical formulations or compositions of the present invention are topical as transdermal salves including various dosage forms; eg, ointments, creams, topical liquids, gels, sprays, or depot formulations (including small pills). Orally; as tablets, capsules, sugar or film coated tablets, solutions, or suspensions orally: via the rectum as suppositories; for example, parenterally, such as intramuscular or intravenous injection or infusion Can be administered. According to a preferred embodiment, the compounds of the invention are designed for topical administration.

  The dosage will depend on age, weight, patient health, and mode of administration; for example, a suitable dosage for oral administration in adults is from about 10 to about 150-1000 mg per dose per day. 1 to 5 times.

  The present invention includes a pharmaceutical formulation or composition comprising a compound of the present invention together with a pharmaceutically acceptable excipient (which is a carrier or diluent).

  The pharmaceutical compositions containing the compounds of the invention are usually prepared according to the following conventional methods and administered in a pharmaceutically suitable form.

  For topical administration, the drug is preferably formulated as an ointment, cream, gel, emulsion or topical solution. Formulation as a powder or oil is also contemplated. The basis of the formulation is familiar to those skilled in the cosmetic and pharmaceutical industry and need not be described in detail here. For example, vegetable oils and lipids such as almond oil, peanut oil, olive oil, peach core oil, sprinkle oil, plant extracts, essential oils; vegetable waxes and synthetic and animal oils, fats or waxes; lecithin, lanolin alcohol, Carotenes, fragrances, mono- or polyhydric alcohols, urea, preservatives, colorants and others can be used. Formulation as an oil in water or a water emulsion in oil is preferred.

  A suitable content of the active compound in the drug is that 4-hydroxytestosterone or a salt or ester thereof is 0.0001-20 wt.%, Preferably 0.6-10 wt.%, More preferably 1-5 wt. Also good. Usually, it is 0.6 to 5% by weight.

  When a compound for promoting skin permeability is mixed, the content thereof is, for example, 0.01 to 1% by weight, preferably 0.05 to 0.2% by weight when hyaluronidase is used. If dimethyl sorbitol or DMSO is used, it may be 1 to 25% by weight, preferably 5 to 10% by weight.

  For example, solid oral forms can be used together with the active compound for diluents such as lactose, dextrose, saccharose, cellulose, corn starch, or potato starch; for example, silica, talc, stearic acid, magnesium stearate or calcium stearate A lubricant such as starch, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, or polyvinylpyrrolidone; a dissociator such as starch, alginate, arginine, or sodium glycolate starch; Colorants, sweeteners; wetting agents such as lecithin, polysorbate, lauryl sulfate; and generally non-toxic and pharmaceutically inert substances used in pharmaceutical formulations. The pharmaceutical preparation can be prepared by known methods such as mixing, granulating, tableting, sugar coating or film coating processes. Liquid dispersions for oral administration include, for example, syrups, emulsions, and suspensions.

  The syrup can contain, for example, saccharose or saccharose with glycerin and / or mannitol and / or sorbitol as a carrier.

  Suspensions and emulsions can contain, for example, natural rubber, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose or polyvinyl alcohol as a carrier.

  Suspensions or solutions for intramuscular injection, together with the active compound, for example, sterilized water, olive oil, pharmaceutically acceptable carriers such as ethyl oleate, for example glycols such as propylene glycol, and the like If so, it can contain an appropriate amount of lidocaine hydrochloride.

  Solutions for intravenous injection or infusion can contain, for example, sterile water as a carrier, or preferably the solution can be a sterile, aqueous, isotonic salt solution.

  Suppositories can contain, together with the active compound, pharmaceutically acceptable carriers such as cocoa butter, polyethylene glycol, polyoxyethylene sorbitan fatty acid ester surfactants or lecithin.

  Where the target tumor is estrogen receptor (ER) positive, the compounds of the present invention can be combined with known estrogen receptor antagonists (such as Tamoxifen (R)) and / or known aromatase inhibitors and / or known Thus, the pharmaceutical composition of the present invention can also be administered in known effective amounts (one type of estrogen receptor antagonistic and / or aromatase inhibitory activity and / or cell growth inhibitory activity). According to a preferred embodiment, the combination of 4-hydroxytestosterone and a suitable cell growth inhibitor is in particular in the form of a formulation that can be administered topically (eg cream ).

  Examples of suitable aromatase inhibitors are disclosed, for example, in US Published Patent 2004 / 0018991A1, and formestane and lentalone, exemestane, MDL 18962, 7 alpha-substituted androstenedione derivatives, ATD, 10-oxirane and 10- Steroidal aromatase inhibitors such as thiirane-substituted androgens, atamestan, and others, and non-steroidal aromatase inhibitors such as borazole, arimidex, letrozole, fadrozole and logletimide. See, for example, “Red List”, Editio Cantor, Aulendorf (DE, 1999) for symbolic representation and availability of these compounds.

  Examples of suitable cell growth inhibitors and combinations with cell growth inhibitors include, for example, AC ± T-taxol (chemical name: paclitaxel) or taxotere (chemical name: docetaxol) Adriamycin (Chemical name: doxorubicin) with or without cyclophosphamide (trade name: Cytoxan); CMF-cyclophosphamide, methotrexate and fluorouracil ("5-FU" or 5- CAF-cyclophosphamide, adriamycin and fluorouracil ("5-FU" or 5-fluorouracil); CEF-cyclophosphamide, epirubicin (adriamycin-like) and fluorouracil ("5-FU" or 5-fluorouracil) FAC-fluorouracil ("5-FU" or 5-fluoro Uracil), there is adriamycin and cyclophosphamide.

  In general, particularly when the target tissue is estrogen receptor negative, the compounds of the present invention can also be administered with an antibody against Her2neu, such as, for example, trastuzumab (Herceptin®). The composition can also include an effective amount of a known compound that exhibits Her2neu antagonist activity.

  The following examples illustrate the invention but are not intended to limit the invention.

Example 1
The compound of the present invention can be synthesized as follows.
As a first step, 2.5 g testosterone is dissolved in 100 ml cold MeOH. After adding 9 ml NaOH (2%) and 17 ml H ₂ O ₂ (30%), the mixture is stirred at 4 ° C. for 24 hours. The resulting epoxide is precipitated with ice water.
As a second stage, 2 g of dry epoxide is dissolved in 200 ml of acetic acid containing 2% H ₂ SO ₄ . The solution is stirred at room temperature for 4 hours. The reaction product is precipitated with ice water.
The reaction product is then washed with 1% NaOH solution to hydrolyze the acetyl ester. The overall yield of purified 4-hydroxytestosterone is in the range of 40-50%.

Example 2
The cream for topical administration of the present invention can be formulated in a conventional manner using the following amounts of ingredients. The amount is given per 100 g of cream:
4-hydroxy 17β-acetylandrost-4-en-3-one 4.5 g
Cetearyl alcohol 7.5g
Paraffin wax 3.0g
Sodium carbomer 2.5g
Isopropyl myristate 6.0g
Sorbitan monostearate 1.0g
Polysorbate 20 3.0g
Stearyl alcohol 2.0g
Dimethylsorbitol 5.0g
Pure water qs 100.0g
The resulting cream is applied topically to the skin over the affected breast tissue of a female or male patient.

Example 3
The gel of the present invention can be formulated in a conventional manner using the following amounts of ingredients. The amount is given per 100 g of gel:
4-hydroxytestosterone 2.5g
95 degree ethanol 70.0g
Carbopol 980 0.5g
Isopropyl myristate 2.5g
Triethanolamine 0.5g
Pure water qs 100.0g

Example 4
The solution used as the spray of the present invention can be prepared by a conventional method using the following formulation. The amount is given per 100 g of solution:
4-hydroxytestosterone 2.5g
95 degree ethanol 70.0g
Isopropyl myristate 2.5g
Pure water qs 100.0g

Example 5
Tablets each weighing 0.150 g and containing 25 mg of active compound can be prepared as follows:
Composition (for 10,000 tablets)
250 g of 4,17β-dihydroxyandrost-4-en-3-one
Lactose 800g
Corn starch 415g
Talc powder 30g
Magnesium stearate 5g
The 4,17β-dihydroxyandrost-4-en-3-one, lactose and half of the corn starch are mixed; the mixture is passed through a 0.5 mm mesh size sieve. Corn starch (10 g) is suspended in warm water (90 ml) and granulated with the resulting paste into a powder. The granules are dried and granulated on a 1.4 mm mesh size sieve, after which the remaining amount of starch, talc and magnesium stearate are added, mixed carefully and processed into tablets.

Example 6
Capsules containing 20 mg of active compound can be prepared with each portion at 0.200 g:
Composition for 500 capsules:
4,17β-dihydroxyandrost-4-en-3-one 10 g
Lactose 80g
Corn starch 5g
Magnesium stearate 5g
This formulation is wrapped in two hard gelatin capsules, the amount for each capsule being 0.200 g.

Example 7
MCF-7 cells were cultured in RPMI 1640 medium at 37 ° C. until the cell number was about 1 × 10 ⁵ cells / ml. Thereafter, four sets of the same MCF-7 cell samples were dihydrotestosterone (DHT) or 4-hydroxytestosterone (4-OHT) to different concentrations, ie 10 ⁻⁷ M, 10 ⁻⁸ M, or 10 ⁻⁹ M. In addition, cell proliferation was measured by measuring viable cells. Cell numbers were counted after 3, 6 and 9 days during incubation at 37 ° C. As a control, the proliferation of MCF-7 cells was measured without the addition of active reagent.
Inhibition of MCF-7 cell proliferation by DHT or 4-OHT compound concentrations of 10 ⁻⁷ M, 10 ⁻⁸ M, and 10 ⁻⁹ M, the respective concentrations are shown in FIGS. 1A, 1B and 1C, and control Compared with. These results show that when using the compound 4-hydroxytestosterone of the present invention, MCF-7 cell proliferation is significantly stronger than 4-dihydrotestosterone and especially stronger than the control, which promotes a significant inhibitory effect. Indicated.
The surprising stimulatory effect compared to DHT is that the effect of this compound is that 4-OHT can bind specifically and strongly to the androgen receptor (AR) and also exhibits an anabolic / apoptotic effect rather than an androgenic effect. Can be explained. In contrast, DHT exhibits an androgenic effect in particular.

Example 8
A double-blind clinical trial was conducted on a group of 10 female patients. Of particular note in this clinical trial is the hormone receptor status of progesterone receptor (PR) and in particular estrogen receptor (ER) of the patient's target breast cancer tissue; Androgen receptor (AR) positive was assumed. In this group, 6 patients had histologically malignant breast cancer (C. mammae). The five tumors in this investigated subgroup were found to be progesterone receptor (PR) negative and in particular estrogen receptor (ER) negative (state level 0 (zero)). For Her-2-neu, 3 patients showed a negative condition, one with moderate (level 1) and the other with strong (level 2) receptor status. In the sixth patient, malignant breast cancer was confirmed, but the amount of cancerous tissue was too small to measure receptor status.
The remaining 4 patients showed no malignant disease but appeared to have normal or mastitic breast tissue.
All 10 female patients were treated with 4-hydroxytestosterone as the active substance, topically and partially, applying the cream described in Example 2 to the target intermammary skin. After continuous local administration every day for 3 months, clinical examination for therapeutic effect was performed.

The results of 6 patients with malignant breast cancer are shown in Table 1 below, along with the receptor status of the first through fifth patients.

Of the women who underwent post-treatment examination, three showed significant recovery of breast cancer tissue as measured by mammography.
Although the tumor receptor status of the sixth patient in this cancer subgroup is unknown, a clear clinical recovery was confirmed in this patient as well.
It should be noted that for the remaining 4 patients with non-malignant disease state, there is a clear reduction in tissue that appears to be normal or mastopathy in the course of local administration.
As a result of these clinical studies, two major important effects of the pharmaceutical composition according to the present invention, including 4-hydroxytestosterone (4-OHT), were found. First, 4-OHT has an excellent effect in shrinking cancer tissue, and second, and surprisingly, even ER negative cancer cells can be effectively treated.
In summary, the findings made in the present invention support the notion that 4-OHT works effectively via AR and that apoptosis can be realized as a result.
Thereby, the discovery of the present invention allows for effective therapeutic and / or prophylactic treatment. Among other useful applications, including adjuvant therapy, neoadjuvant therapy is particularly important by using a significant reduction in tumor volume to assist or enable subsequent surgical treatment planning.

Example 9
A 42 year old patient with bilateral breast cancer and skin metastasis was treated daily with the cream according to Example 2. After 4 weeks of treatment, the skin metastases disappeared.

Example 10
A 50 year old patient with large breast cancer (diameter: 12 cm), huge tumor spread and metastatic liver disease was subjected to partial tumor resection, followed by daily treatment with the cream according to Example 2. During the treatment, the patient showed no further tumor growth and even a reduction of the unresectable tumor mass.

Example 11
A 74 year old patient with advanced breast cancer and tumor expansion to lymph nodes was treated daily for 10 weeks. A significant decrease in tumor volume occurred, and the affected lymphocytes no longer showed a tumor.

The compound concentration of DHT or 4-OHT shows an inhibition results of MCF-7 cell growth when the ^{10 -7} M. It is a figure which shows the inhibition result of MCF-7 cell proliferation when the compound density | concentration of DHT or 4-OHT is made into 10 ^<-8 > M. It is a figure which shows the inhibition result of MCF-7 cell proliferation when the compound density | concentration of DHT or 4-OHT is 10 ^<-9 > M.

Claims (8)

Breast cancer (particularly breast mucinous ductal carcinoma, medullary carcinoma of the breast and breast lobular carcinoma), endometrial cancer, cervical cancer of 4,17β-dihydroxyandrost-4-en-3-one or a salt or ester thereof , Epithelial ovarian cancer, uterine (fallopian tube) cancer, benign proliferative change or alteration of the breast, fibroadenoma, micromastopathy (perimenopause), mastopathy, uterine fibroids, and secondary or metastases from these diseases Use in the preparation of a pharmaceutical composition for prevention or treatment of sexual tumors. Breast cancer (especially breast mucinous ductal carcinoma, medullary and breast lobular carcinoma), endometrial cancer, cervical cancer, epithelial ovarian cancer, uterine (fallopian tube) cancer, benign proliferative change or alteration of the breast 4,17β-dihydroxyandrost-4-en-3- 3 for the prevention or treatment of fibroadenoma, fibroadenoma (small menopause), mastopathy, hysteromyoma, and secondary or metastatic tumors derived from these diseases ON or a salt or ester thereof. Breast cancer (especially breast mucinous ductal carcinoma, medullary and breast lobular carcinoma), endometrial cancer, cervical cancer, epithelial ovarian cancer, uterine (fallopian tube) cancer, benign proliferative change or alteration of the breast A method for the prevention or treatment of fibroadenoma, micromastopathy (perimenopause), mastopathy, uterine fibroids, and secondary or metastatic tumors derived from these diseases, for patients in need of such treatment Or a method of prevention or treatment comprising administering an effective amount of 4,17β-dihydroxyandrost-4-en-3-one. 4,17β-dihydroxyandrost-4-en-3-one is an effective amount of one or more estrogen receptor antagonists and / or one or more aromatase inhibitors and / or one or more cell growth inhibitors 4. The method of claim 3, wherein the method is administered with an agent and / or one or more antibodies against Her2neu. The method according to claim 3 or 4, wherein the administration is local. A pharmaceutical composition for the prevention and / or treatment of female or male patients comprising 4,17β-dihydroxyandrost-4-en-3-one as active ingredient and comprising a suitable carrier and / or diluent . 7. An effective amount of one or more estrogen receptor antagonists and / or one or more aromatase inhibitors and / or one or more cytostatics and / or one or more antibodies against Her2neu. A pharmaceutical composition according to 1. 8. A pharmaceutical composition according to claim 6 or 7 formulated for topical administration.

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